PMID:10386615
| Citation |
Takagi, H, Kobayashi, C, Kobayashi, S and Nakamori, S (1999) PCR random mutagenesis into Escherichia coli serine acetyltransferase: isolation of the mutant enzymes that cause overproduction of L-cysteine and L-cystine due to the desensitization to feedback inhibition. FEBS Lett. 452:323-7 |
|---|---|
| Abstract |
PCR random mutagenesis in the cysE gene encoding Escherichia coli serine acetyltransferase was employed to isolate the mutant enzymes that, due to a much less feedback inhibition by L-cysteine, cause overproduction of L-cysteine and L-cystine in the recombinant strains. The L-cysteine auxotrophic and non-utilizing E. coli strain was transformed with plasmids having the altered cysE genes. Then, several transformants overproducing L-cysteine were selected by detecting the halo formation of the L-cysteine auxotroph. The production test of amino acids and analysis of the catalytic property on the mutant enzymes suggest that the carboxy-terminal region of serine acetyltransferase plays an important role in the desensitization to feedback inhibition and the high level production of L-cysteine and L-cystine. |
| Links | |
| Keywords |
Acetyltransferases/chemistry; Acetyltransferases/genetics; Acetyltransferases/metabolism; Amino Acid Substitution; Cysteine/biosynthesis; Cystine/biosynthesis; DNA Primers; Escherichia coli/enzymology; Escherichia coli/genetics; Feedback; Kinetics; Mutagenesis, Site-Directed; Point Mutation; Polymerase Chain Reaction/methods; Recombinant Proteins/chemistry; Recombinant Proteins/metabolism; Serine O-Acetyltransferase |
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